BS2092: Common Techniques In Molecular And Cell Biology-Practical- Medical Science Assignment Help

Assignment Task

Question 1.

A. The gel below represents the result of an experiment to purify GST or a GST fusion protein (GST- PLCg-SH2 domain) from bacterial lysates expressing these proteins. Samples are run on a gel and lanes are labelled: A, B, C, D.

Which of the following descriptions corresponds to the sample loaded in lanes A, B, C, D? Highlight the correct answer.

A common vector used for generating GST-fusion proteins is indicated below.

B. Briefly describe how IPTG induces expression of GST fusion proteins cloned into pGEX3X

C. What is the purpose of the ampicillin resistance gene in pGEX3X

Question 2

A. Starting with a liquid culture of bacteria transformed with pGEX3x, or pGEX3X- PLCg-SH2 domain, using bullet points, outline the experimental steps required to generate the gel shown in part A. You do not need to include details such as volumes or concentrations.

B. From the gel shown in question 1A, estimate the size of the PLCg-SH2 domain. Indicate your reasoning.

Question 3

A typical extraction buffer used to lyse the bacteria is: 50mM Tris HCl pH 7.5, 150mM NaCl, 2mM EDTA, 5mM Benzamidine, 1% Triton X-100. You need to make up ONE LITRE of extraction buffer starting with solid chemicals or stock solutions, as shown below. To make this solution you will need to weight out an appropriate mass of solid Benzamidine and NaCl, dissolve these in a small amount of water then add an appropriate volume of Tris HCl solution, EDTA solution and 10% Triton. Then make up the solution to a final volume of 1 L. Protease inhibitor would be added immediately before use.

1. What mass of benzamidine would you need to weight out? Show workings.
2. What volume of 1M Tris HCl, pH7.5 solution would you need to add? Show workings.
3. What volume of 10% Triton X-100 would you add?
4. Why would you use Triton X-100 in the lysis buffer and not SDS?
5. What is the purpose of adding protease inhibitors?

Question 4

A. The following images of a cell in interphase and a cell undergoing mitosis were obtained using immunofluorescence microscopy. A cell in interphase is labelled. In In which phase of the cell cycle is the mitotic cell.

Question 5

1. Green Fluorescent Protein (GFP) can be used instead of antibodies to determine the localization of a protein. Explain in your own words how you would use GFP to visualize the microtubules in a cell.
2. List TWO advantages and TWO disadvantages of using a GFP-fusion protein over immunofluorescence microscopy.

This BS2092 - Medical Science has been solved by our Phd Experts at My Uni Paper. Our Assignment Writing Experts are efficient to provide a fresh solution to this question. We are serving more than 10000+ Students in Australia, the UK and the US by helping them to score HD in their academics. Our Experts are well trained to follow all marking rubrics referencing Style.

Be it a used or new solution, the quality of the work submitted by our assignment experts remains unhampered. You may continue to expect the same or even better quality with the used and new assignment solution files respectively. One thing to be noticed is that you could choose between the two and acquire an HD either way. You could choose a new assignment solution file to get yourself an exclusive, plagiarism (with free Turn tin file), expert quality assignment or order an old solution file that was considered worthy of the highest distinction.